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Abstract: TH-PO1062

Podocyte Protective Effects of TRPC5 Inhibitor AC1903 in Human iPSC-Derived Podocytes and Kidney Organoids

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Zhou, Yiming, Brigham and Women's Hospital/Harvard Medical School, Boston, Massachusetts, United States
  • Emani, Maheswarareddy, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, United States
  • Pablo, Juan lorenzo B., Broad Institute of MIT and Harvard, Cambridge, Massachusetts, United States
  • Greka, Anna, Brigham and Women's Hospital/Harvard Medical School, Boston, Massachusetts, United States
Background

The TRPC5-Rac1 pathway has been shown to induce podocyte injury and loss in two pre-clinical rat models: a transgenic rat FSGS model and a spontaneous hypertensive rat model. The TRPC5 blocker AC1903 protects podocytes in both models. However, less is known about the effect of this compound in rat models that are more easily amenable to drug development programs, such as the puromycin aminonucleoside (PAN)-induced nephrosis model. Furthermore, the effect of AC1903 in human podocytes remains elusive. Here, we aim to investigate the effect of AC1903 in the puromycin aminonucleoside(PAN)-induced nephrosis in rat, as well as in PAN-treated human iPSC-derived podocytes and kidney organoids.

Methods

A single i.p. injection of PAN (50mg/kg) was given to wild-type Sprague-Dawley rats (Male, 4-5 weeks, Charles River). AC1903 was administered twice a day for 7 days after PAN injection. 24-hour urine albumin levels were measured on day 7.
Human iPS cells were used to generate podocytes and kidney organoids (according to Subramanian A. et al. 2019 http://dx.doi.org/10.1101/516807). PAN treatment was used to induce human podocyte injury in these in vitro model systems, and the effects of AC1903 were assayed by Western Blotting, immunofluorescence staining and confocal microscopy.

Results

We found that a single i.p. injection of PAN-induced podocyte injury and foot process effacement (FPE) as well as a significant increase in urine albumin levels 7 days after injection. Treatment of proteinuric PAN rats with AC1903 significantly reduced foot process effacement and proteinuria. PAN treatment of human iPSC-derived podocytes and kidney organoids triggered the TRPC5-Rac1 injury pathway leading to ROS production and cytoskeletal dysregulation. These effects were reversed by AC1903, showing for the first time that TRPC5 inhibition benefits human podocytes and kidney organoids.

Conclusion

Taken together, our results confirmed the relevance of the TRPC5-Rac1 pathway in human kidney tissue thus highlighting the potential of this therapeutic strategy for patients.

Funding

  • NIDDK Support